PEG Adjustment Enables Genome Transplantation Using Mycoplasma Mycoides Recipient

Abstract

Pioneering advances in synthetic biology, initiated at the J. Craig Venter Institute, have enabled the creation of the first cell driven by synthetic genome and opened a new era of creating designer microbes. This technology offers far greater potential for genetic modification than traditional genome editing techniques and holds promise for the routine creation of synthetic cells as DNA synthesis costs decrease and genetic knowledge expands. Two essential technologies underpin this achievement: the ability to clone entire genomes in host organisms, such as Saccharomyces cerevisiae, and the successful transplant of these genomes into recipient cells to generate living organisms. In all previous work the recipient cell in genome transplantation experiments has been Mycoplasma capricolum. In this study, we explored the potential of using Mycoplasma mycoides strains as recipient cells for genome transplantation. By increasing polyethylene glycol (PEG) concentration from 5% to 10%, we successfully transplanted various M. mycoides strains using several M. mycoides recipient cells. Additionally, we demonstrated the ability to transplant M. capricolum genomes into M. mycoides recipient cells; although with lower efficiency compared to M. mycoides strains. These findings provide a modified transplantation protocol and likely get us closer to expanding genome transplantation to other bacterial species.